AGENDA

TOPIC OUTLINE
PRODUCTION OF
MICROBIAL INTRODUCTION 01
BIOASSAY OF BIOLOGICAL

INSECTICIDES ALTERNATIVES TO INSECTICIDES 05

02
CHEMICAL INSECTICIDES
FORMULATION AND USE OF 06
PRESENTATION BY: BIOINSECTICIDES
BIOLOGICAL CONTROL OF INSECTS 03
CALATA & MACADANGDANG SAFETY TESTING OF 07
PRODUCTION OF BIOLOGICAL BIOINSECTICIDES
INSECTICIDES 04

Date: November 06, 2025

INTRODUCTION ALTERNATIVES TO CHEMICAL INSECTICIDES

The alternatives to the use of chemicals include the following:

Insects are major pests of crops. Enormous losses occur when they attack various 1. Predators
plant parts, often transmitting disease in the process. Even after harvest insects Among vertebrates one of the best known is the use of fish especially
attack stored foods; this attack of stored foods is not limited to plant foods, but also Gambusia affinis to eat mosquito larvae. Invertebrate predators include other
includes animal foods such as dried fish. Besides the loss they cause in agriculture larger insects e.g. wasps, while plant predators include Utricularia (a bladder wort).
and food, insects are also vectors of various animal and human diseases.

In recent years, insect control has shifted from relying solely on chemicals to using
integrated methods. This change is due to issues like chemical insecticides
harming both pests and their natural predators, growing resistance, environmental
and health concerns, and rising costs linked to petroleum-based products.

2. Genetic manipulations: 3. The use of hormones or hormone analogs: Pheromones are synthetic
These include the production compounds which act as sex attractants. The insects attracted are
(by chemicals or by destroyed.

irradiation) of large numbers of

4. The use of pathogens: Pathogens of insects are found among bacteria,
sterile males, whose mating fungi, protozoa, viruses and nematodes. The idea of using pathogens to
does not result in control insects originated from studies of the diseases of the silkworm
Bombyx mori. The pioneer work of Bassi was followed by those of Le Conte,
fertile eggs.
Pasteur, Hagen until Metchnikoff actually tested the control of sugar beet
pests with the fungus Metarrhizium anisopliae in South Russia.
 Table 17.1
BIOLOGICAL CONTROL Biological control agents as ranked in order of priority by the special
program of the World Health Organization 1980
OF INSECTS
Biological control is widely applied in agriculture, food
production, and forestry, but less so in controlling disease-
carrying insects like mosquitoes.

In 1976, the World Bank, UNDP, and WHO launched a Special

Programme for Research and Training in Tropical Diseases,
targeting six diseases, four of which are insect-borne:
malaria, trypanosomiasis, filariasis, and leishmaniasis.

The WHO ranked biological control organisms (e.g., bacteria,

fungi, nematodes, fish) for effectiveness against insect
vectors like mosquitoes and black flies.

Table 17.1
Biological control agents as ranked in order of priority by the special
program of the World Health Organization 1980
Table 17.2 Pathogenic viruses found in insects

DESIRABLE PROPERTIES IN ORGANISMS TO BE

Table 17.2 Pathogenic viruses found in insects USED FOR BIOLOGICAL CONTROL

The following are desirable in microorganisms to be used

in the biological control of insect:

(a) The agent should be highly virulent for the target insect, but should kill no other
insects.
(b) The killing should be done quickly so that in the case of crops, damage is kept as
low as possible, and in the case of vectors of disease before extensive transmission
of the disease occurs.
(c) The killing ability should be predictable.
(d) The agent should not be harmful to man, animals or crops; in other words it should
be safe to use.
DESIRABLE PROPERTIES IN ORGANISMS TO BE CANDIDATES WHICH HAVE BEEN CONSIDERED AS
USED FOR BIOLOGICAL CONTROL BIOLOGICAL CONTROL AGENTS

I. Bacteria as Biological Control Agent

The following are desirable in microorganisms to be used
General Traits: Spore-forming bacteria preferred for durability and mass production.
in the biological control of insect:
Key Species
(e) It should be technically amenable to cheap industrial production.
Bacillus thuringiensis (Bt):
(f) When produced, it should be stable under the conditions of use such as under the
(commonly known as ‘Bt’) is an insecticidal bacterium, marketed worldwide for control of
high temperature and ultra violet light of ordinary sunlight. many important plant pests–mainly caterpillars of the Lepidoptera (butterflies and moths) but
(g) It should be viable over reasonably long periods to permit storage and also mosquito larvae, and simuliid blackflies that vector river blindness in Africa.
transportation as necessary.
(h) It should ideally persist or recycle and/or be able to search for its host. Bacillus moritai: This is used in Japan for the same purpose as B. thuringiensis
serotypes H3 and H3A.
Bacillus popilliae: Obligate pathogen of Japanese beetle; produced in beetle
larvae.
Bacillus sphaericus: Targets mosquito larvae.

Bacillus popilliae: : This is an obligate pathogen of the Japanese beetle II. Viruses as Biological Control Agents
Popilla japonica against which it is used. Since it is an obligate parasite it is
produced in the larvae of the beetle. A large number of viruses has been isolated from insects. The advantages of
viruses as biological control agents is that they are specific.
Bacillus thuringinensis var. israelensis (also known as serotype H14). This
was isolated in 1976 by Goldberg and Margalit from a mosquito breeding Seven groups of insect pathogentic viruses have been identified (Table 17.2).
site in Israel. It has proved very effective in killing mosquito larvae and the The most useful of them for biological control purposes are the baculoviruses,
black fly (Simulium spp). which are easily recognizable because the virus particles are included within a
Bacillus sphericus: Bacillus sphericus is an highly specific for mosquito proteinaceous inclusion body large enough to be seen under a light microscope.
larvae as Bacillus thuringiensis, var israelensis (B.t.i.).

II. Viruses as Biological Control Agents III. Fungi as Biological Control Agent
The baculoviruses are the best candidates for insect control because All the four major groups of fungi, Phycomycetes, Ascomycetes, Fungi
they are: Imperfecti and Basidiomycetes contain members pathogenic to insects.

(a) effective in controlling insect populations, The great difficulty with using fungi for biological control is that
(b) restricted to a host range of invertebrates, environmental conditions including temperature and humidity must be
(c) relatively easy to produce in large quantities and adequate for spore germination and insect cuticle penetration by the
(d) stable under specific conditions because of the inclusion bodies. hyphae. Since these environmental conditions are not always assured the
result is that fungi are used for biological control only in a few countries
especially the USSR.
III. Fungi as Biological Control Agent IV. Protozoa as Biological Control Agent

. Fungi which have been most widely used as Beauvaria bassiana and In constrast to the rapid action of viruses and spore-forming bacteria,
Metarrhizium anisopliae. Others are Hirsutella thompsonii Verticillium and killing by protozoa is slow and may take weeks. Furthermore they are
Aschersonia aleyrodis. H. thompsonii is being developed commercially as difficult to produce, being accomplished only in vivo. Nevertheless
acaricide, for killing mites which attack plants, although a large number of they have been produced and successfully used experimentally for
other fungi attack mites. H. thomposonii has been found particularly active stored-product pests (Matosia trogoderina) mosquitoes (Nosema
against mites which attack citrus. algerae) and grasshoppers (Nosema pyrasta).

IV. Protozoa as Biological Control Agent

Vavra vilivis is also effective against mosquitoes and has properties
similar to those of Nogema algerae. Studies sponsored by the WHO
have shown that N. algerae does not seem to constitute a safety PRODUCTION OF
hazard for man. Factors favoring the use of N. algerae are spore
longevity, ease of spore-production under laboratory and especially
BIOLOGICAL INSECTICIDES
cottage industry conditions and the probable impact on disease
transmission by reducing the longevity of infected female
mosquitoes.

MICROBIOLOGICAL INSECTICIDE PRODUCTION METHODS

PRODUCTION OF BIOLOGICAL I. SUBMERGED FERMENTATION

INSECTICIDES These have been used for the production of Bacillus spp. (excluding production of
B. poppillae which is produced in vivo) and to a lesser extent, fungi.
Microbiological insecticides are produced in one of three ways:
submerged fermentation; surface or semi-solid fermentation;
Medium: In fermentation for Bacillus thuringiensis the active principle sought is
and in vivo production. The first two are for facultative
the delta toxin found in the crystals. Media for submerged fermentation have
pathogens and the third is for obligate pathogens.
been compounded by various workers in a number of patents. In one such
preparation, the initial growth in a shake flask occurred in nutrient broth; in the
second shake flask, and in the seed fermentor best molasses (1%), corn steep
liquour (0.85%) and CaCO3 (0.1%) were used.
 MICROBIOLOGICAL INSECTICIDE PRODUCTION METHODS MICROBIOLOGICAL INSECTICIDE PRODUCTION METHODS

I. SUBMERGED FERMENTATION II. SURFACE CULTURE

Bacillus thuringiensis var insraelensis and Bacillus sphericus do not require
Surface culture techniques are used for fungi and for spore formers. The
carbohydrates for growth and can grow well and produce materials which will
organisms after shake-flask growth are cultured in a seed tank from where the
kill the larvae of mosquitoes in a variety of proteinaceous materials such as
broth is transferred to flat bins with perforated bottoms.
commercial powders of soy products, dried milk products, blood and even
The semi-solid medium is a mixture of an agricultural by product such as bran,
materials from primary sewage tanks. Effective powders of B. sphericus 1593
an inert product such as kisselghur, soy bean meal, dextrose, and mineral salts.
and B. thuringiensis var israelensis have been produced using discarded cow
The use of this medium increases the surface area and hence aeration because
blood from abattoirs and various legumes.
of the thinness of its spread in the bins.
Hot air is passed through the perforations to dry the material. It is ground,
Extraction: At the end of the fermentation, the active components of the broth
assayed and compounded to any required strength with inert material.
are recovered by centrifugation, vacuum filtration with filter aid or by
Submerged, culture in which the hyphae are used have been carried out with
precipitation. Precipitation has been done with CaCl2 and the acetone
good results in the United States using Hirsutella thompsonii.
method yields products of very high potency. Thefermentation beer may
readily be diluted and used directly.

MICROBIOLOGICAL INSECTICIDE PRODUCTION METHODS

III. In vivo production

In vivo culture methods are used for producing caterpillar viruses, mosquito protozoa
and Bacillus popillae.
The method is labor-intensive and could be easily applied for suitable candidates in
developing countries where expertise for submerged culture production is usually
lacking.
BIOASSAY OF
Once the organism has been obtained in a sufficient quantity to last for several years it
is lyophilized and stored at low temperature.
The viruses are introduced into the food of the larvae and the dead larvae are crushed,
BIOLOGICAL
centrifuged to remove large particles and the rest are dried.
The amount of viruses in each larva is variable but the virus content of between one
INSECTICIDES
and one hundred caterpillars should be sufficient to treat one acre in the case of cotton
moths.
Usually separate facilities are used for rearing the caterpillars, for infecting them and
for the extraction of the virus particles. The preparation is then bio assayed and mixed
with a suitable carrier.

BIOASSAY OF BIOLOGICAL FORMULATION AND USE OF

AGENDA
INSECTICIDES
AGENDA
BIOINSECTICIDES

Standards do exist for Bacillus thuringiensis serotypes H3 and H3A The formulation of the bioinsecticides is extremely important. An insecticide
used against caterpillars and a standard for B. thuringiensis var shown to be highly potent under laboratory experimental conditions may
israelensis against mosquito ‘IPS82’ exists. prove valueless in the field unless the formulation has been correctly done.
Both standards are prepared and deposited at the Institute Pasteur in Since microorganisms cannot by themselves be patented, industrial firms
Paris. In the simplest terms a standard is based on the LD50, the dose of producing bioinsecticides depend for their profits on the efficiency of their
the insecticide which will kill 50% of the population must be clearly formulation (i.e., the inert material which ensures adequate presentation of
defined; the age and type of insect to be used; the food of the insect; the larvicide to the target insect).
the temperature conditions and a host of other parameters.
 FORMULATION AND USE OF FORMULATION AND USE OF
AGENDA
BIOINSECTICIDES
AGENDA
BIOINSECTICIDES

Dusts
The inert material is referred to as a carrier or an extender. Carriers or Semi-solid preparations based on waste plant products usually are compounded
extenders are the solids or liquids in which the active principle is diluted. When as dusts or powders because making them into liquid causes the bran to absorb
the carrier is a liquid and the active principle is suitable in it the application is water and prevent free flow thus leading to the clogging of conventional liquid
a spray. There are thus two types of formulation: applicators.
The advantage of dusts is greater stability of the preparation.
(a) powders and dusts They are also useful when the insecticide is intended to reach the underside of low
(b) flowable liquid; which of the two is manufactured depends to a large extent lying crops such as cabbages.
Heavy rains unfortunately wash off dusts. They may also lead to inhalation of the
on the method of production and intended use of the insecticide.
bioinsecticides by the persons applying them.

FORMULATION AND USE OF

AGENDA
BIOINSECTICIDES

Liquid Formulation
Liquid formulations are usually made from water in which both the crystal and
SAFETY TESTING OF
spores are stable. Sometimes oils and water/oil emulsions may be used. When BIOINSECTICIDES
liquids other than water are used it must be ascertained that they do not
inactivate the active agent. Emulsifiers may be added to stabilize emulsions
when these are used.

SAFETY TESTING OF
AGENDA
BIOINSECTICIDES

Public Concern: Use of microorganisms in pest control often raises safety fears.
Testing Methods: SEARCH AND
Animal tests include: oral ingestion, inhalation, intraperitoneal, intradermal,
intravenous inoculation.
DEVELOPMENT OF NEW
Additional tests: teratogenicity (birth defects) and carcinogenicity (cancer risk).
Test Subjects: Rats, mice, monkeys, rabbits, fish, and occasionally human volunteers.
BIOINSECTICIDES
Proven Safe (Non-toxic):
Bacteria: Bacillus popillae, B. thuringiensis, B. moritai
Viruses: Heliothus, Orgyia, Lymantria, Autographa, Dendrolimus
Protozoa: Nosema locustae, N. algerae, N. troqodermae
Fungi: Beauveria bassiana, Hirsutella thompsonii
 SEARCH AND DEVELOPMENT SEARCH AND DEVELOPMENT
AGENDA
OF NEW BIOINSECTICIDES
AGENDA
OF NEW BIOINSECTICIDES

WHO Guidelines: Follows a structured scheme for screening, safety,

evaluation, and environmental impact (Table 17.3). Timeline:
Stages of Development: Typically 5–7 years to develop a new bioinsecticide.
Discovery and screening of entomopathogens. May be faster (<5 years) if safety data exists for a related organism.
Safety and environmental testing. Cost Advantage:
Formulation and field trials. Development cost is 20–50% lower than that of chemical insecticides.
Industrial production and distribution.
Production Scale:
Small-scale (cottage industry) possible for some agents.
Most require industrial expertise for formulation and marketing.

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